Cervical cancer is the leading cause of cancer-related death in women. It causes over 250,000 death world-wide / year, with 80% occurring in countries without systematic cervical neoplasia screening programs5. High risk Human Papillomavirus (HPV) infection is responsible for >99% of all the cervical cancers4. Early detection of HPV infection by the Pap Test has reduced the death toll in developed nations significantly10. Despite its merits, the Pap Test is no longer considered an adequate screening technology due to the high false-negative and false-positive rates as a result of subjective visual interpretation of cell morphology by pathologists7. In recent years, several nucleic acid-based HPV diagnostic tests have emerged8. These tests have excellent clinical sensitivity, but fail to provide the desired clinical specificity, due to the high prevalence of HPV infection without pathological outcome8. Consequently, utilization of molecular HPV tests for screening may lead to costly over-treatment of patients (follow-up visits; colposcopies; biopsies). In response to the need for a specific yet sensitive screening method, Arbor Vita Corporation proposes the development of a diagnostic test that detects the Papillomavirus protein E6, a pivotal oncoprotein in cervical oncogenesis. E6 oncoprotein levels in cervical swab specimens from patients with neoplasia correlate positively with the severity of lesions6. Accordingly, an HPV-E6 oncoprotein-based screening test has the potential to identify HPV-positive women who are at risk to develop cervical cancer amidst the much larger number of women with clinically irrelevant HPV infection. In this grant, the applicants propose the development of a high-throughput bead-based assay to detect and to type E6 oncoproteins from 15 high-risk HPV types for use in centralized clinical labs. The new HPV-E6 oncoprotein test will harness the technological advances in bead-based immunoassays and the availability of highly specific HPV-E6 monoclonal antibodies at Arbor Vita to achieve the desired high sensitivity, specificity and capability for automation. Under phase 1, our approach is focused on the development of the bead-based 15 HPV type E6 oncoprotein diagnostic prototype test. Upon successful completion of phase 1, the applicants will submit a phase 2 grant application for further product development towards the final format of the test that will be used in clinical studies for design validation. PUBLIC HEALTH RELEVANCE: Cervical cancer is caused by HPV and is worldwide a leading cause of death in women; implementation of sensitive and specific screening technology is the key to reduced mortality. The Pap Test screening method is no longer considered adequate. The applicants propose the development of an HPV-E6 oncoprotein based test for cervical neoplasia screening. Via detection of E6, the cancer causing agent, the technology promises to identify those women who need clinical attention, among many more women who have HPV infection without clinical outcome. This should translate to public heath cost-saving by reducing the number of unnecessary clinical follow-up and treatment.